TY - JOUR
T1 - Mechanisms of DNA uptake by mammalian cells
T2 - Fate of exogenously added DNA monitored by the use of fluorescent dyes
AU - Loyter, A.
AU - Scangos, G. A.
AU - Ruddle, F. H.
PY - 1982
Y1 - 1982
N2 - Coprecipitation of DNA with calcium phosphate is a commonly used method of gene transfer in mammalian cells. We have found that DNA forms a tight complex with Ca P[i] and that DNA in this complex is resistant to nucleases present in serum or added externally. Under optimal conditions, virtually all of the recipient mouse Ltk-Aprt- cells take up Ca P[i]-DNA complexes, as determined by fluorescent dyes specific for DNA (4',6-diaminilo-2-phenylindol dihydrochloride) or for calcium salts (chlorotetracycline). However, only a small proportion of the cells have detectable Ca P[i]-DNA complexes in the nucleus. Uptake of the CaP[i]-DNA complexes was highly dependent upon the pH at which the CaP[i]-DNA complex was formed and upon the concentration of DNA in the complex.
AB - Coprecipitation of DNA with calcium phosphate is a commonly used method of gene transfer in mammalian cells. We have found that DNA forms a tight complex with Ca P[i] and that DNA in this complex is resistant to nucleases present in serum or added externally. Under optimal conditions, virtually all of the recipient mouse Ltk-Aprt- cells take up Ca P[i]-DNA complexes, as determined by fluorescent dyes specific for DNA (4',6-diaminilo-2-phenylindol dihydrochloride) or for calcium salts (chlorotetracycline). However, only a small proportion of the cells have detectable Ca P[i]-DNA complexes in the nucleus. Uptake of the CaP[i]-DNA complexes was highly dependent upon the pH at which the CaP[i]-DNA complex was formed and upon the concentration of DNA in the complex.
UR - http://www.scopus.com/inward/record.url?scp=0006500895&partnerID=8YFLogxK
U2 - 10.1073/pnas.79.2.422
DO - 10.1073/pnas.79.2.422
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C2 - 6952193
AN - SCOPUS:0006500895
SN - 0027-8424
VL - 79
SP - 422
EP - 426
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 2 I
ER -